解剖学报

解剖学报 ›› 2013, Vol. 44 ›› Issue (2 ): 204-209.doi: 10.3969/j.issn.0529-1356.2013.02.011

• 细胞和分子生物学 • 上一篇    下一篇

乳酸抑制脂多糖介导的核因子-κB p65入核转录及环氧化酶-2转录

姜金奇1 许光勇1  史雅然1  乔雨1  胡格1,2  任晓明1*   

  1. 1. 北京农学院动物科技学院，北京 102206; 2.北京农学院兽医学(中医药)北京市重点实验室，北京 102206
  • 收稿日期:2012-06-18 修回日期:2012-08-23 出版日期:2013-04-06 发布日期:2013-04-06
  • 通讯作者: 任晓明 E-mail:13910224882@163.com
  • 基金资助:

    北京市自然科学基金（6072007）；北京市教委资助项目（KM200910020002）;省自然科学基金资助项目;自然科学基金资助项目

Lactic acid inhibits lipopolysaccharide-induced translocation of NF-κB p65 from cytoplasm to nucleus and transcription of nuclear factor-κB p65 and cyclooxygenase 2

JIANG Jin-qi1 XU Guang-yong1 SHI Ya-ran1 QIAO Yu 1 HU Ge 1,2 REN

Xiao-min 1*
  

  1. 1．Department of Animal Science and Technology，Beijing University of Agriculture，Beijing 102206，China;2．Beijing Key Laboratory of Traditional Chinese Veterinary Medicine，Beijing University of Agriculture，Beijing 102206，China
  • Received:2012-06-18 Revised:2012-08-23 Online:2013-04-06 Published:2013-04-06

摘要：

目的 探讨嗜酸乳杆菌代谢产物乳酸(LA）对内毒素（LPS）介导的NF-κB p65入核、转录及环氧化酶-2（COX-2）转录的抑制作用。方法 以体外培养的大鼠肠黏膜微血管内皮细胞（RIMMVECs）为材料，设对照组、LPS组、LA预处理组和NF-κB特异性阻断剂吡咯烷二硫代氨基甲酸盐（PDTC）预处理组4个实验组。LPS作用30min后，Western blotting 检测不同实验组细胞核、细胞质内NF-κB p65蛋白水平，LPS作用9h后，实时荧光定量PCR法检测NF-κB p65、COX-2 mRNA水平。结果 LPS作用30min后，各实验组细胞NF-κB p65蛋白总量差异不显著，但LA和PDTC预处理组细胞核内NF-κB p65比例显著低于LPS组；LPS作用9h后，LA和PDTC预处理组细胞NF-κB p65和COX-2 mRNA水平显著低于LPS组。结论 乳酸具有类似NF-κB阻断剂的作用，可以抑制LPS介导的NF-κB p65入核、转录，抑制NF-κB下游靶基因如COX-2的转录，从而发挥抗炎作用。

关键词: 乳酸 , 脂多糖 , 核因子-κB , 环氧化酶-2 , 微蛋白内皮细胞 , 免疫印迹法;实时荧光定量聚合酶链反应 , 大鼠

Abstract:

Objective To investigate whether lactic acid(LA),which was extracted from the culture supernatant of Lactobacillus,could inhibit lipopolysaccharide(LPS)-induced translocation of NF-κB p65 from cytoplasm to nucleus and transcription of NF-κB p65 and cyclooxygenase 2(COX-2). Methods Rat intestinal mucosa microvascular endothelial cells(RIMMVEs) were cultured in vitro and were divided into 4 groups:control group,LPS group,LA pretreated group and NF-κB inhibitor (pyrrolidinecarbodithioate,PDTC) pretreated group.The protein of NF-κB p65 in cytoplasm and in nucleus was detected by Western blotting analysis after RIMMVEs were treated with LPS for 30 minutes.The mRNA of NF-κB p65 and COX-2 was detected by real time quantity-PCR after RIMMVEs were treated with LPS for 9 hours. Results In a short time(treated with LPS for 30 minutes),the protein expression of NF-κB p65 had no notable difference but the ratios of p65 in nuclei of LA and PDTC pretreated group were notably lower than that of LPS group.After a longer time(treated with LPS for 9 hours),the mRNA expression of NF-κB p65 and COX-2 were notably lower than LPS group. Conclusion LA has the effects like NF-κB inhibitor,notably inhibiting LPS-induced translocation of NF-κB p65 from cytoplasm to nucleus and transcription of NF-κB p65 and COX-2.By this way,LA inhibits inflammatory response.

Key words: Lactic acid , Lipopolysaccharide , Nuclear factor-κB , Cyclooxygenase 2 , Mucosa microvascular endothelial cells, Western blotting, Real time quantity-PCR , Rat